Protein Native gels

Biochemistry Enzymology
1

Hi ,

I trying to run a purified 29 KDa protein in Native gels, but always it is stuck in the wells ( when I’ m not using stackig gel) or at the beguining of the separating gel. This protein is a dimer with a pI of 8.3 , i have tried to run it in native gels with pH 9.3 but it doesn’ t work, it is still stuck in the wells. Does anybody know what could be the problem?

2

Laura

Have you tried using a running buffer with alternative pH? If not, my suggestion is to use a running buffer with a pH close to your protein’s pI (around pH 8.0-8.5). Also, ensure the gel concentration is appropriate for the size of your protein (~8-12% acrylamide).